The overall objective of our project is the development of the first 3D in vitro platform able to screen efficiently and reliably the health and nutritional value of sustainable feeds that the industry is currently developing.
This will be achieved through these intermediate objectives:
- The development of a functional fish artificial intestine (Fish-AI) able to mimic the complex functions of the intestinal mucosa. The prototype will combine a purposely-developed 3-D scaffold, with the growth and differentiation of a functional intestinal epithelium. Scaffold permeability to nutrients, its ability to promote cell proliferation and differentiation will be measured to define success (month 24).
- The assessment and development of the Fish-AI prototype capability to predict the nutritional and health value of feeds and bioactive components. Morphological, biochemical, cellular and molecular data in response to a well-characterized gut health challenger (soybean meal) will be collected. These data will be contrasted with classical fish functional parameters detected in response to the administration of the same meals in an in vivo Univariate and multivariate analyses coupled with canonical correlation and a principal component analysis will enable us to pinpoint the parameters that define the reliability and power of prediction of the Fish-AI platform (month 36).
- The validation of Fish-AI through the analysis of previously untested fish feeds. The nutrition and health value of two feeds never tested before, will be evaluated through the parallel use of Fish-AI and in vivo trials as described above. The degree of correlation between in vivo and in vitro data will give the measure to which degree the platform can provide a reliable prediction of the functional response of a complex organ (month 48).
The following science-to-technology breakthroughs will be targeted by the project:
- Identification of a material with the correct stiffness and, at the same time, allowing cell attachment, growth and differentiation.
- The material must also ensure a physiological permeability to biological molecules like amino acids, sugars and lipids after these are absorbed by the overlaying epithelial monolayer.
- Techniques must be developed for moulding such material with sufficient resolution to mimic the microscopic architecture of the native fish gut.
- Biomaterials and culture techniques must take into account that the body temperature of the model species (Rainbow trout) is below 20°C instead of the typical 37°C commonly used with mammalian cells.
- Appropriate and effective source of intestinal epithelial cells must be found for the derivation of cell lines that can generate a heterogeneous population that recreates the complex structure of the intestinal mucosa but allows to obtain repeatable results.
Scientific and technological contributions to the foundation of a new future technology
The wide employment of the Fish-AI platform will provide a wealth of biological data that are currently too difficult to obtain through whole-organism experiments due to their inherent complexity. This next generation in vitro platform will enable to combine the complexity of a 3-dimensional heterogeneous tissue-like population with the ease of access and analysis of traditional in vitro culture models.
The public is increasingly uneasy with the use of animal models, irrespectively of how good the reason for their usage is. The innovative 3D in vitro technology that will result from this project has the ambition to constitute a substantial leap towards a reliable and relevant alternative to the use of animal models. In addition to reducing the number of animals used, the platform will also ensure better animal welfare. Compared to in vivo trials in fact, the use of Fish-AI will reduce the time needed to screen and develop new raw materials, thus speeding the elimination of all harmful feed components.
Another area where Fish-AI could reduce the use of animal models is neonatology and premature infant nutrition that currently adopt new-born piglets as model of choice for mimicking pathologies such as short bowel syndrome and for the development of new baby formulas. An appropriate artificial intestine, in fact, would enable the direct use of human cells with obvious beneficial outcomes in terms of ease of clinical translation and a substantial reduction of animal experiments.
Potential for future social or economic impact or market creation
European aquaculture is a recognized leader in technological development and know-how creation. Farming of salmonids is considered the most sophisticated of its kind, and developments for these species are often transferred and adapted to other species, farmed in developing Countries. If effective, the Fish-AI could be easily adapted to other species, therefore the presence of a global company in the consortium may facilitate its use to benefit the food production from aquaculture in other countries too.
Building leading research and innovation capacity across Europe.
The consortium will expose several Early Stage Researchers to different research and working settings ranging from traditional academic institutions to high-tech SME through the R&D division of a large industrial company. The high degree of interdisciplinarity of the consortium, together with the intense exchange program of material, data and personnel will provide the opportunity for high level training of these young members of the teams. They will form a new cohort of trained and skilled individuals with a high degree of employability within aquaculture and beyond, most of whom are first-time participants to FET under Horizon 2020.
Other foreseen substantial impacts that would address issues related to climate change and environment include the possible use of the Fish-AI platform to address the ecological impact of traditional and alternative feed sources. The technologies that will be developed by the project, in fact, will provide an ideal tool for the study of effluent discharge in aquatic environment and could be applied not only to aquaculture and mariculture but to terrestrial intensive farming as well.
A fully functional Fish-AI would pave the way for a wide array of innovative products based on 3D models of complex organs creating new markets opportunities that strengthen the competitiveness and growth of active and ambitious high-tech SME or academic spin-offs.